摘要
为了探讨荧光显微技术在酿酒酵母细胞不同研究方面的作用,通过GFP标记目标蛋白的同源重组的方法和免疫荧光技术标记两种蛋白,最后利用荧光显微镜观察酵母细胞中某种蛋白定位及两种蛋白共定位情况;分别用荧光染液DAPI、FM4-64、BODIPY、Filipin、DHE和Annexin V试剂处理酵母细胞之后,利用荧光显微镜观察细胞中的细胞核、液泡、脂滴、麦角固醇、ROS和细胞凋亡的情况。结果显示,荧光显微技术在酵母细胞蛋白定位、细胞器观察及细胞中ROS和细胞凋亡等研究方面具有重要作用。
To investigate the role of fluorescence microscopy in different studies of S. cerevisiae cells, the method of labeling homologous recombination of the target protein by GFP and the immunofluorescence technique were used to label the two proteins. Finally, the protein localization and co-localization of two proteins in S. cerevisiae were observed by fluorescence microscopy. After the treatment of S. cerevisiae with fluorescent dye DAPI,FM4-64, BODIPY, Filipin, DHE and Annexin V, the nucleus, vacuoles, liposomes, ergosterol, ROS and cell apoptosis were observed by fluorescence microscopy. The results showed that fluorescence microscopy could play an important role in the research of protein localization, organelle observation, ROS and apoptosis in S. cerevisiae cells.
作者
曹春蕾
曹正锋
赵运英
Cao Chunlei;Cao Zhengfeng;Zhao Yunying(National Engineering Laboratory for Cereal Fermentations Technology, Jiangnan University, Wuxi, 214122;College of Animal Science and Tech- nology, Yangzhou University, Yangzhou, 225009)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2018年第4期1539-1546,共8页
Genomics and Applied Biology
基金
国家自然科学基金(31301021)资助
关键词
荧光显微技术
酿酒酵母
蛋白定位
免疫荧光
细胞器染色
Fluorescence microscopy
Saccharomyces cerevisiae
Protein localization
Immunofluorescence
Organelle staining